Genetic Engineering Publications - GEG Tech top picks
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Programmable RNA editing by recruiting endogenous ADAR using engineered RNAs - Nature Biotech

Programmable RNA editing by recruiting endogenous ADAR using engineered RNAs - Nature Biotech | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
Cellular RNAs are edited with high specificity using engineered ADAR-recruiting RNAs.
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In this article, the scientists  present an approach, called leveraging endogenous ADAR for programmable editing of RNA (LEAPER), that employs short engineered ADAR-recruiting RNAs (arRNAs) to recruit native ADAR1 or ADAR2 enzymes to change a specific adenosine to inosine. They pave the way for a single-molecule system, LEAPER, enabling precise, efficient RNA editing with broad applicability for therapy and basic research.

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Unbiased detection of off-target cleavage by CRISPR-Cas9 and TALENs using integrase-defective lentiviral vectors - Nature Biotechnology

Unbiased detection of off-target cleavage by CRISPR-Cas9 and TALENs using integrase-defective lentiviral vectors - Nature Biotechnology | Genetic Engineering Publications - GEG Tech top picks | Scoop.it

The utility of CRISPR-Cas9 and TALENs for genome editing may be compromised by their off-target activity. We show that integrase-defective lentiviral vectors (IDLVs) can detect such off-target cleavage with a frequency as low as 1%. In the case of Cas9, we find frequent off-target sites with a one-base bulge or up to 13 mismatches between the single guide RNA (sgRNA) and its genomic target, which refines sgRNA design.



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 Several groups have reported that the linear double-stranded IDLV genome could be incorporated preferentially into DNA double-strand breaks (DSBs) by nonhomologous end-joining (NHEJ). In this study, the scientists report the use of IDLVs to measure the off-target activities of CRISPR-Cas9 and TALENs. These results suggest that TALEN and CRISPR–D10A Cas9 (a mutant  nuclease with nickase activity), have improved target specificity over CRISPR-Cas9.


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